Generation of Induced Pluripotent Stem Cells Using Recombinant Proteins

Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Cell stem cell (Impact Factor: 22.15). 05/2009; 4(5):381-4. DOI: 10.1016/j.stem.2009.04.005
Source: PubMed


Available from: Gary Siuzdak, Jan 16, 2014
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    ABSTRACT: Induced Pluripotent Stem Cells (iPSCs) are self renewable and can differentiate to different types of adult cells, which has shown great promises in the field of regenerative medicine. iPSCs are reprogrammed from human somatic cells through ectopic expression of various transcription factors viz. Oct4, Sox2, Klf4, and c-Myc (OSKM). This novel technology enables derivation of patient specific cells, which possess a potential cure for many diseases. During the last decade, significant progresses have been achieved in enhancing the reprogramming efficiency, safety of iPSCs derivation, development of different delivery techniques by various research groups. Nevertheless, it is important to resolve and define the mechanism underlying the pluripotent stem cells. Major bottleneck which arises during iPSCs generation is the availability of source material (cells/tissues), difficulty to deliver transcription factors with no aberrant genetic modifications and limited reprogramming efficiency. Reprogramming may be achieved by employing different cocktails with number of different transcription factors, application of miRNA and some small molecules such as (Valproic acid, CHiR99021, Sodium butyrate, Vitamin C, Parnate etc). Similarly, various starting source materials have been demonstrated for iPSC based therapies including fibroblasts, cord blood, peripheral blood, keritinocytes, urine, etc., with their specific uses and limitations. Moreover, with the advent of many new reprogramming techniques, various direct delivery methods have been introduced such as using synthetic mRNA expressing pluripotent gene network has been shown to be an appropriate technique to deliver transcription factors and a dozen of small molecules which can replace transcription factors or enhance reprogramming efficiency. This article addresses the iPSCs technology mechanisms, progresses and current perspectives in the field.
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    ABSTRACT: Direct reprogramming technology has emerged as an outstanding technique for the generation of induced pluripotent stem (iPS) cells and various specialized cells directly from somatic cells of different species. Recent studies dissecting the molecular mechanisms of reprogramming have methodologically improved the quality, ease and efficiency of reprogramming and eliminated the need for genome modifications with integrating viral vectors. With these advancements, direct reprogramming technology has moved closer to clinical application. Here, we provide a comprehensive overview of the cutting-edge findings regarding distinct barriers of reprogramming to pluripotency, strategies to enhance reprogramming efficiency, and chemical reprogramming as one of the non-integrating approaches in iPS cell generation. In addition to direct transdifferentiation, pluripotency factor-induced transdifferentiation or cell activation and signaling directed (CASD) lineage conversion is described as a robust strategy for the generation of both tissue-specific progenitors and clinically relevant cell types. Then, we consider the possibility that a combined method of inhibition of roadblocks (e.g. p53, p21, p57, Mbd3, etc.), and application of enhancing factors in a chemical reprogramming paradigm would be a safe, reliable and effective approach in pluripotent reprogramming and transdifferentiation. Furthermore, with respect to the state of native, aberrant, and target gene regulatory networks in reprogrammed cell populations, CellNet is reviewed as a computational platform capable of evaluating the fidelity of reprogramming methods and refining current engineering strategies. Ultimately, we conclude that a faithful, highly efficient and integration-free reprogramming paradigm would provide powerful tools for research studies, drug-based induced regeneration, cell transplantation therapies and other regenerative medicine purposes.
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    ABSTRACT: Ever since a technology to reprogram somatic cells into pluripotent stem cells was developed by Dr. Shinya Yamanaka’s group in 2006, its therapeutic potential has been extensively discussed. We now call the reprogrammed embryonic stem cell (ESC)-like cell an ‘induced pluripotent stem cell’ (iPSC). The beauty and power of the iPS in human case is that it avoids many ethical issues, in that unlike human ESCs (hESCs), iPSCs do not require destroying a human embryo to establish pluripotent cell lines. The iPSC holds many hopes that many human diseases may be treatable in the near future. On the other hand, there are still several issues that need to be solved prior to the therapeutic use of iPSCs in humans directly. The biggest hurdle is that, so far, there is lack of ways to completely exclude tumorigenic iPSC-derived cells. Additionally, there is an issue that immune rejection may occur, even in autologously grafted iPSCs, as was observed in monkey experiment. Nevertheless, iPSCs, combined with genetic manipulation, hold much promise that iPSCs may be used in cell therapy procedures and as tools for investigating underlying mechanisms of human diseases. This review will discuss recent progress in cellular reprogramming and its potential use in regenerative medicine.
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